Traditional chinese medicine composition and use thereof in treatment of dementia

Abstract

A traditional Chinese medicine composition for treating dementia, containing: radix hedysari, Chinese skullcap, wild Asparagus, and Chuanxiong rhizome, and further containing velvet antler and / or monkshood.

Description

Traditional Chinese medicine composition and its use in treating dementia Technical Field

[0001] The present invention relates to a Chinese medicine composition comprising: Astragalus root, Scutellaria root, Asparagus cochinchinensis root, and Ligusticum chuanxiong. The present invention also relates to the use of the Chinese medicine composition in treating dementia. Background Art

[0002] Dementia can be categorized by cause into degenerative dementia (caused by neurodegeneration), vascular dementia (caused by cerebral vascular blockage or disease), and mixed dementia (a combination of these two causes). Among these dementias, Alzheimer's disease (AD) is the most common.

[0003] Although reports have shown that cholinesterase inhibitors can be used to slow the progression of dementia, their effectiveness is limited and they have many undesirable side effects. Therefore, there is currently no clinically effective cure for dementia.

[0004] Therefore, finding drugs from natural sources that can effectively treat dementia is the goal that researchers in this field are committed to. Summary of the Invention

[0005] Through research, the applicant unexpectedly discovered that a traditional Chinese medicine composition containing Astragalus root, Scutellaria root, Asparagus cochinchinensis root, and Ligusticum chuanxiong rhizome not only effectively treats Alzheimer's disease (AD), but also improves spatial memory and promotes neurogenesis. Furthermore, the composition is able to treat ischemia-reperfusion-induced nerve injury, suggesting high potential for the treatment of dementia.

[0006] Therefore, in a first aspect, the present invention provides a traditional Chinese medicine composition comprising: Astragalus membranaceus, Scutellaria baicalensis, Asparagus cochinchinensis, and Ligusticum chuanxiong.

[0007] In a second aspect, the present invention provides a Chinese medicine composition for use in preparing a medicine for treating dementia, wherein the Chinese medicine composition comprises: Astragalus membranaceus, Scutellaria baicalensis, Asparagus cochinchinensis, and Ligusticum chuanxiong.

[0008] In a third aspect, the present invention provides a method for treating dementia, comprising administering a Chinese medicine composition to a subject in need thereof, wherein the Chinese medicine composition comprises: Astragalus membranaceus, Scutellaria baicalensis, Asparagus cochinchinensis, and Ligusticum chuanxiong.

[0009] In a fourth aspect, the present invention provides a traditional Chinese medicine composition for treating dementia, comprising: Astragalus membranaceus, Scutellaria baicalensis, Asparagus cochinchinensis, and Ligusticum chuanxiong.

[0010] In a specific embodiment, the traditional Chinese medicine composition comprises: 7-11 parts by weight of Astragalus membranaceus, 2-6 parts by weight of Scutellaria baicalensis, 4-7 parts by weight of Asparagus cochinchinensis, and 2-4 parts by weight of Chuanxiong rhizome. In an exemplary embodiment, the traditional Chinese medicine composition comprises: 8-10 parts by weight of Astragalus membranaceus, 3-5 parts by weight of Scutellaria baicalensis, 5-6 parts by weight of Asparagus cochinchinensis, and 3 parts by weight of Chuanxiong rhizome.

[0011] According to the present invention, the traditional Chinese medicine composition may further include deer antler. In a specific embodiment, the traditional Chinese medicine composition includes: 7-11 parts by weight of Astragalus membranaceus, 2-6 parts by weight of Scutellaria baicalensis, 4-7 parts by weight of Asparagus cochinchinensis, 2-4 parts by weight of Chuanxiong rhizome, and 1-3 parts by weight of deer antler. In an exemplary embodiment, the traditional Chinese medicine composition includes: 8-10 parts by weight of Astragalus membranaceus, 3-5 parts by weight of Scutellaria baicalensis, 5-6 parts by weight of Asparagus cochinchinensis, 3 parts by weight of Chuanxiong rhizome, and 2 parts by weight of deer antler.

[0012] According to the present invention, in addition to the above five ingredients, the traditional Chinese medicine composition may further include aconite root. In a specific embodiment, the traditional Chinese medicine composition includes: 7-11 parts by weight of Astragalus membranaceus, 2-6 parts by weight of Scutellaria baicalensis, 4-7 parts by weight of Asparagus cochinchinensis, 2-4 parts by weight of Chuanxiong rhizome, 1-3 parts by weight of antler, and 1-3 parts by weight of aconite root. In an exemplary embodiment, the traditional Chinese medicine composition includes: 8-10 parts by weight of Astragalus membranaceus, 3-5 parts by weight of Scutellaria baicalensis, 5-6 parts by weight of Asparagus cochinchinensis, 3 parts by weight of Chuanxiong rhizome, 2 parts by weight of aconite root, and 2 parts by weight of antler.

[0013] According to the present invention, the dementia is selected from the group consisting of degenerative dementia, vascular dementia, and mixed dementia. In a specific embodiment, the dementia is a degenerative dementia selected from the group consisting of Alzheimer's disease (AD), dementia with Lewy bodies (DLB), frontal-temporal dementia (FTD), Parkinson's disease dementia (PDD), and Huntington's disease (HD). In an exemplary embodiment, the dementia is Alzheimer's disease.

[0014] According to the present invention, the treatment of dementia includes promoting neurogenesis. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] FIG1 shows the discrimination index measured for each group of mice in Example 2, wherein “*”, “**”, and “***” indicate p < 0.05, p < 0.01, and p < 0.001, respectively;

[0016] FIG2 shows the elution profile obtained by subjecting the Chinese medicine composition 3 of the present invention to high performance liquid chromatography;

[0017] FIG3 shows the digging weights measured for each group of mice in Example 4, where “*” and “**” indicate p < 0.05 and p < 0.01, respectively;

[0018] FIG4 shows the nesting scores of mice in each group measured in Example 4, wherein “***” indicates p < 0.001;

[0019] FIG5 shows the escape latency measured for each group of mice in Example 4, where “*” indicates p < 0.05;

[0020] FIG6 shows the number of Aβ plaques measured in the brains of mice in each group in Example 4, where “***” indicates p<0.001;

[0021] FIG7 shows the soluble Aβ40 levels measured in the cerebral cortex of each group of mice in Example 4, where “*” and “***” indicate p<0.05 and p<0.001, respectively;

[0022] FIG8 shows the escape latency measured for each group of mice in Example 5, where “*” and “**” indicate p < 0.05 and p < 0.01, respectively;

[0023] FIG9 shows the number of Aβ plaques measured in the brains of mice in each group in Example 5, where “*” and “***” indicate p<0.05 and p<0.001, respectively;

[0024] FIG10 shows the DCX measured in the subgranular area of ​​the dentate gyrus of the brain of each group of mice in Example 5. + Cell number, where “*” and “**” indicate p < 0.05 and p < 0.01, respectively; and

[0025] FIG11 shows the results of immunofluorescence staining analysis of the hippocampus of the brains of each group of mice in Example 7. DETAILED DESCRIPTION

[0026] For the purposes of this specification, it will be expressly understood that the word "comprising" means "including, but not limited to," and that the word "comprises" has a corresponding meaning.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein that can be used to practice the present invention. Of course, the present invention is in no way limited to the methods and materials described.

[0028] The present invention provides a traditional Chinese medicine composition comprising: Astragalus membranaceus, Scutellaria baicalensis, Asparagus cochinchinensis, and Chuanxiong rhizome. In a specific embodiment, the traditional Chinese medicine composition comprises: 7-11 parts by weight of Astragalus membranaceus, 2-6 parts by weight of Scutellaria baicalensis, 4-7 parts by weight of Asparagus cochinchinensis, and 2-4 parts by weight of Chuanxiong rhizome. In an exemplary embodiment, the traditional Chinese medicine composition comprises: 8-10 parts by weight of Astragalus membranaceus, 3-5 parts by weight of Scutellaria baicalensis, 5-6 parts by weight of Asparagus cochinchinensis, and 3 parts by weight of Chuanxiong rhizome.

[0029] According to the present invention, the traditional Chinese medicine composition may further include deer antler. In a specific embodiment, the traditional Chinese medicine composition includes: 7-11 parts by weight of Astragalus membranaceus, 2-6 parts by weight of Scutellaria baicalensis, 4-7 parts by weight of Asparagus cochinchinensis, 2-4 parts by weight of Chuanxiong rhizome, and 1-3 parts by weight of deer antler. In an exemplary embodiment, the traditional Chinese medicine composition includes: 8-10 parts by weight of Astragalus membranaceus, 3-5 parts by weight of Scutellaria baicalensis, 5-6 parts by weight of Asparagus cochinchinensis, 3 parts by weight of Chuanxiong rhizome, and 2 parts by weight of deer antler.

[0030] According to the present invention, in addition to the above five ingredients, the traditional Chinese medicine composition may further include aconite root. In a specific embodiment, the traditional Chinese medicine composition includes: 7-11 parts by weight of Astragalus membranaceus, 2-6 parts by weight of Scutellaria baicalensis, 4-7 parts by weight of Asparagus cochinchinensis, 2-4 parts by weight of Chuanxiong rhizome, 1-3 parts by weight of antler, and 1-3 parts by weight of aconite root. In an exemplary embodiment, the traditional Chinese medicine composition includes: 8-10 parts by weight of Astragalus membranaceus, 3-5 parts by weight of Scutellaria baicalensis, 5-6 parts by weight of Asparagus cochinchinensis, 3 parts by weight of Chuanxiong rhizome, 2 parts by weight of aconite root, and 2 parts by weight of antler.

[0031] According to the present invention, the red astragalus root, scutellaria root, asparagus root, ligusticum chuanxiong, pilose antler, and aconite root in the Chinese medicinal composition can be fresh traditional Chinese medicinal materials, or products obtained by a processing selected from the group consisting of drying, grinding, chopping, comminuting, processing, and combinations thereof. In a specific embodiment, the aconite root in the Chinese medicinal composition has been processed to remove toxicity, and is also known as Zhi Fu Zi or Paw Fuh Tzyy.

[0032] According to the present invention, the red scutellaria in the traditional Chinese medicine composition can be derived from the dried root of Hedysarum polybotrys Hand.-Mazz. The scutellaria in the traditional Chinese medicine composition can be derived from the dried root of Scutellaria baicalensis Georgi. The asparagus in the traditional Chinese medicine composition can be derived from the dried root of Asparagus cochinchinensis (Lour.) Merr. The chuanxiong in the traditional Chinese medicine composition can be derived from the dried rhizome of Ligusticum chuanxiong Hort. The velvet antler in the traditional Chinese medicine composition can be derived from the dried young antlers of species in the Cervidae family (Alces, Axis, Cervus, Elaphurus, Rangifer, or Rusa), particularly from the dried young antlers of red deer (Cervus elaphus Linnaeus), sika deer (Cervus nippon taiouanus), or sambar deer (Rusa unicolor swinhoii). In an exemplary embodiment, the velvet antler is derived from the dried young antlers of red deer. The aconite root in the traditional Chinese medicine composition can be derived from the dried root of Aconitum carmichaelii Debeaux.

[0033] According to the present invention, the Chinese medicine composition can be manufactured into a dosage form suitable for oral administration using techniques well known to those skilled in the art. According to the present invention, dosage forms suitable for oral administration include, but are not limited to: commercially available traditional Chinese medicine powders, or traditional Chinese medicine liquids or traditional Chinese medicine extracts obtained by decocting Chinese medicinal materials, and capsules and other preparations further prepared from the above-mentioned traditional Chinese medicine powders, liquids or extracts. In a specific embodiment, the Chinese medicine composition is prepared by mixing traditional Chinese medicine powders or extracts of Astragalus membranaceus, Scutellaria baicalensis, Asparagus cochinchinensis, Chuanxiong, Pilose Antler and Aconite.

[0034] According to the present invention, the Chinese medicinal materials (i.e., Astragalus, Scutellaria, Asparagus cochinchinensis, Chuanxiong, Pilose Antler and Aconite) used to prepare the Chinese medicine composition can be decocted together or separately. In a specific embodiment, the Chinese medicine composition is obtained by mixing the Chinese medicinal materials and then decocting them in water. According to the present invention, the weight ratio of the Chinese medicinal materials to water may fall within the range of 1:8 to 1:25. In a specific embodiment, the weight ratio of the Chinese medicinal materials to water falls within the range of 1:10 to 1:12. In an exemplary embodiment, the weight ratio of the Chinese medicinal materials to water is 1:10. According to the present invention, the decoction temperature may fall within the range of 70 to 100°C. In an exemplary embodiment, the decoction temperature is 100°C. According to the present invention, the decoction time may fall within the range of 30 to 120 minutes. In a specific embodiment, the decoction time falls within the range of 30 to 90 minutes. In an exemplary embodiment, the cooking time is 60 minutes.

[0035] In addition, the present invention provides a Chinese medicine composition for treating dementia, wherein the Chinese medicine composition is as described above. The present invention also provides a use of the Chinese medicine composition as described above for preparing a medicine for treating dementia.

[0036] As used herein, "treating" or "treatment" means preventing, reducing, alleviating, ameliorating, relieving, or controlling one or more clinical signs of a disease or disorder, as well as lowering, stopping, or reversing the progression of the severity of a condition or symptom being treated.

[0037] According to the present invention, the treatment of dementia includes promoting neurogenesis. As used herein, the terms "neurogenesis" and "neuroregeneration" are used interchangeably.

[0038] According to the present invention, the dementia is selected from the group consisting of degenerative dementia, vascular dementia, and mixed dementia. In a specific embodiment, the dementia is a degenerative dementia selected from the group consisting of Alzheimer's disease (AD), dementia with Lewy bodies (DLB), frontal-temporal dementia (FTD), Parkinson's disease dementia (PDD), and Huntington's disease (HD). In an exemplary embodiment, the dementia is Alzheimer's disease.

[0039] According to the present invention, the medicine may further comprise a pharmaceutically acceptable carrier widely used in drug manufacturing technology. For example, the pharmaceutically acceptable carrier may comprise one or more agents selected from the group consisting of a solvent, a buffer, an emulsifier, a suspending agent, a decomposer, a disintegrating agent, a dispersing agent, a binding agent, an excipient, a stabilizing agent, a chelating agent, a diluent, a gelling agent, a preservative, a wetting agent, a lubricant, an absorption delaying agent, a liposome, a taste masking agent, an absorption enhancer, a sustained-release agent, a molding agent, an adhesive, a nutritional additive, a colorant, and the like. The selection and amount of these agents are within the professional knowledge and routine skills of those skilled in the art.

[0040] According to the present invention, the pharmaceutical product can be manufactured into a dosage form suitable for oral administration using techniques well known to those skilled in the art, including, but not limited to, sterile powders, tablets, troches, lozenges, pellets, pills, capsules, powders, dispersible powders or granules, solutions, suspensions, emulsions, syrups, elixirs, slurries, pastes for internal use, and the like.

[0041] The present invention also provides a method for treating dementia, comprising administering the above-mentioned Chinese medicine composition to a subject in need thereof.

[0042] As used herein, the terms "administering" and "administration" are used interchangeably and mean introducing, providing or delivering a predetermined active ingredient to a subject by any appropriate route to perform its intended effect.

[0043] As used herein, the term "subject" means any mammal of interest, such as humans, monkeys, cows, sheep, horses, pigs, goats, dogs, cats, mice, and rats.

[0044] According to the present invention, the dosage and frequency of administration of the pharmaceutical product will vary depending on the severity of the disease to be treated, the route of administration, and the age, physical condition and response of the individual to be treated. Generally speaking, the pharmaceutical product can be in the form of a single dose or divided into several doses.

[0045] <Example>

[0046] General experimental materials:

[0047] 1. Experimental Animals

[0048] [Corrected 02.04.2025 according to Rule 26] Male APP / PS1 mice (4-5 months old, weighing approximately 30 g) and male C57 / BL6 mice (4-5 months old, weighing approximately 25 g) used in the following examples were purchased from the Laboratory Animal Center in Taiwan, China, and male ICR mice (adult, weighing approximately 28-30 g) were purchased from BioLasco Taiwan Co., Ltd. All experimental animals were individually housed in a vivarium with a 12-hour light and dark cycle, a room temperature maintained at 22±1°C, and a relative humidity maintained at 65±5%, and adequate water and feed were provided.

[0049] General experimental methods:

[0050] 1. Statistical analysis:

[0051] In the following examples, experiments were repeated three times for each group, and the experimental data are expressed as mean ± standard deviation (SD). One-way analysis of variance (ANOVA) was performed, followed by Tukey's honest significant difference test as a post hoc test to assess differences between the groups. A statistically significant result of p < 0.05 was considered.

[0052] Example 1. Preparation of the Chinese medicine composition of the present invention

[0053] The Chinese medicinal materials were mixed according to Table 1 below, and the resulting mixture was then immersed in 1000 mL of hot water for 30 minutes. The mixture was then boiled at 100°C for approximately 60 minutes until approximately 300 mL of liquid remained. The mixture was then filtered through filter paper (No. 1) and the filtrate collected. The collected filtrate was then freeze-dried to obtain Chinese medicinal compositions 1 to 3 of the present invention.

[0054] Table 1. Formulas of Chinese medicine compositions 1 to 3 of the present invention

[0055] Example 2. Effect of the Chinese herbal composition of the present invention on the recognition memory function of mice with Alzheimer's disease (AD)

[0056] Experimental methods:

[0057] A. Administration of the Chinese medicine composition of the present invention:

[0058] First, APP / PS1 mice (5 months old) were randomly divided into one pathological control group and three experimental groups (i.e., experimental groups 1 to 3), and C57 / BL6 mice (5 months old) served as a normal control group, with n = 5 per group. The mice in experimental groups 1 to 3 were administered with the Chinese medicinal compositions 1 to 3 prepared in Example 1 (all at a dose of 100 mg / kg, in normal saline) via oral gavage, while the pathological control group and the normal control group were administered with an equal amount of normal saline once daily for a total of 5 weeks. At the end of the 5th week after the start of dosing, the mice in each group were subjected to the following experiment in Section B.

[0059] B. Novel object recognition test:

[0060] This test was conducted in an open box that had been cleaned with ethanol to remove odors, over a total of three days. On day one, each group of mice was placed in the center of the open box for 5 to 10 minutes to allow the mice to acclimate to the environment. On day two, each group of mice was placed in the center of the open box, and two identical objects were placed in two diagonal corners for the mice to explore for 10 minutes. On day three, each group of mice was placed in the center of the open box, and one of the two objects was replaced with a different object (i.e., a new object and an old object were placed in the two diagonal corners, respectively) for the mice to explore for 5 minutes. The time the mice spent exploring the new and old objects was recorded. "Object exploration" is defined as the mouse approaching an object and making physical contact with it with its snout and / or forepaw.

[0061] The discrimination index is calculated by substituting the measured exploration time of the new object and the old object into the following formula (1):

[0062] Formula (1): A = B / (B + C)

[0063] Where: A = discrimination index

[0064] B = Time spent exploring new objects

[0065] C = Time to explore old objects

[0066] The obtained experimental data were then analyzed according to the method described in item 1 of the “General Experimental Methods” above.

[0067] result:

[0068] A. New object recognition test:

[0069] Figure 1 shows the discrimination index measured for each group of mice. As shown in Figure 1, the discrimination index of mice in the pathological control group was significantly reduced compared to the normal control group, indicating that the APP / PS1 mice had developed AD and had developed defects in their ability to recognize novel objects. Furthermore, the discrimination index of mice in experimental groups 1 through 3 was significantly increased compared to the pathological control group. These experimental results demonstrate that Chinese herbal compositions 1 through 3 of the present invention can improve recognition and memory function in mice with AD, demonstrating their high potential for treating AD.

[0070] Next, the applicant selected Chinese medicine composition 3 as a representative to conduct the following experiments.

[0071] Example 3. High performance liquid chromatography (HPLC) analysis of the Chinese medicine composition of the present invention

[0072] Experimental methods:

[0073] The Chinese medicine composition 3 of the present invention was dissolved in ddH2O to obtain a mixture with a final concentration of 50 mg / mL, and then the mixture was filtered using a filter membrane with a pore size of 0.22 μm. The filtrate was then collected as a test sample and subjected to HPLC analysis. The HPLC analysis instrument used was as follows: SHIMADZU LC-2050C HPLC system; the analytical column was C18-AR-IIODS column, length: 250 mm x 4.6 mm. The operating conditions for HPLC are shown in Table 2 below.

[0074] Table 2. HPLC operating conditions

[0075] result:

[0076] Figure 2 shows the elution profile of the present invention's Chinese herbal composition 3 subjected to high performance liquid chromatography. As shown in Figure 2, the present invention's Chinese herbal composition 3 exhibits numerous elution peaks during the retention period from 0 to 85 minutes, indicating a rather complex composition.

[0077] Example 4. Evaluation of the therapeutic effect of the Chinese herbal composition of the present invention on mice with AD

[0078] Experimental methods:

[0079] A. Administration of the Chinese medicine composition of the present invention:

[0080] First, APP / PS1 mice (5 months old) were randomly divided into 1 pathological control group and 1 experimental group, and C57 / BL6 mice (5 months old) were used as the normal control group, with n=5 in each group. The experimental group mice were administered with the Chinese medicine composition 3 prepared in Example 1 (at a dose of 100 mg / kg in normal saline) by oral gavage, while the pathological control group and the normal control group mice were administered with an equal amount of normal saline, administered once a day for a total of 8 weeks. At the end of the 5th week after the start of administration, each group of mice was taken for the following tests of B and C, at the end of the 6th week after the start of administration, each group of mice was taken for the following test of D, and at the end of the 8th week after the start of administration, each group of mice was sacrificed and the brain was removed, and then the analysis of E and F was performed respectively.

[0081] B. Burrowing test:

[0082] A cylinder containing 230 g of food pellets was placed in the home cage for mice in each group to dig, and the remaining food pellets in the cylinder were weighed after 2 hours.

[0083] The obtained experimental data were then analyzed according to the method described in item 1 of the “General Experimental Methods” above.

[0084] C. Nesting test:

[0085] Two cotton sheets were placed in the cage as nesting material. After 24 hours, the mice in each group were observed to see if they tore up the nesting material and used it to build a nest. The nest integrity was scored on a scale of 1 to 5, with higher scores indicating greater nest integrity.

[0086] The obtained experimental data were then analyzed according to the method described in item 1 of the “General Experimental Methods” above.

[0087] D. Morris water maze test:

[0088] The water maze was set up in a circular pool (120 cm in diameter, 40 cm in depth) with a water level of 20 cm and a water temperature maintained at 22-24°C. The pool was divided into four equal quadrants, and a platform (10 cm in diameter) was placed 1 cm below the water surface in one quadrant.

[0089] First, each group of mice underwent a 6-day training session, with four trials conducted daily, with 20-minute intervals between each trial. In each trial, each group of mice was randomly placed in one of three starting positions in the pool, facing the pool wall. Regardless of whether the mice could find the platform within 60 seconds, they were allowed to stay on the platform for 30 seconds. On the 6th day of testing, a computer video image analysis system was used to record the average time it took the mice to find the platform across the four trials as their escape latency.

[0090] The obtained experimental data were then analyzed according to the method described in item 1 of the “General Experimental Methods” above.

[0091] E. Immunofluorescence staining assay:

[0092] The brains of mice in each group were fixed with 4% formaldehyde at 4°C and then sliced ​​(about 30 μm thick) using a freezing microtome. The obtained tissue sections were stained with anti-Aβ1-16 antibody (AB10, Millipore cat#MAB5208) as the primary antibody and Alexa Fluor 500 antibody (Alexa Fluor 5000). Immunofluorescence staining was performed using 647-conjugated anti-IgG antibody (Jackson ImmunoResearch) as a secondary antibody according to well-known and commonly used techniques in the art. Stained tissue sections were observed and photographed using a confocal fluorescence microscope (Zeiss LSM 780), and the number of Aβ plaques was counted using Image J software.

[0093] The obtained experimental data were then analyzed according to the method described in item 1 of the “General Experimental Methods” above.

[0094] F. Enzyme-linked immunosorbent assay (ELISA):

[0095] The cerebral cortex of each group of mice was placed in a homogenization buffer containing 4% SDS and protease inhibitors to form a mixture, which was then sonicated. The mixture was centrifuged at 100,000 x g for 1 hour at 4°C. The supernatant was collected and neutralized with Tris buffer (1 M, pH 11) to obtain a test sample. The soluble Aβ40 content in the test sample was then measured using an Amyloid beta 40 ELISA kit (Invitrogen, KHB3482) according to the manufacturer's instructions.

[0096] The obtained experimental data were then analyzed according to the method described in item 1 of the “General Experimental Methods” above.

[0097] result:

[0098] A. Excavation test:

[0099] Figure 3 shows the measured digging weights of mice in each group. As shown in Figure 3, the digging weights of mice in the pathological control group were significantly reduced compared to those in the normal control group, indicating that the APP / PS1 mice had developed AD and had developed deficits in their innate behavioral abilities. Furthermore, the digging weights of mice in the experimental group were significantly increased compared to the pathological control group, even approaching those of mice in the normal control group. This experimental result demonstrates that the traditional Chinese medicine composition of the present invention can effectively treat AD and improve the innate behavioral abilities of mice with AD.

[0100] B. Nesting test:

[0101] Figure 4 shows the nesting scores measured for each group of mice. As shown in Figure 4, compared to the normal control group, the nesting scores of the pathological control group mice were significantly reduced, indicating that the APP / PS1 mice had developed AD and had developed deficits in their activities of daily living (ADLs). Furthermore, compared to the pathological control group, the nesting scores of the experimental group mice were significantly increased, even approaching those of the normal control group mice. This experimental result demonstrates that the Chinese herbal composition of the present invention can effectively treat AD and improve the ability of AD-bearing mice to perform ADLs.

[0102] C. Morse water maze test:

[0103] Figure 5 shows the escape latency measured for each group of mice. As shown in Figure 5, the escape latency of the pathological control group mice was significantly increased compared to the normal control group, indicating that the APP / PS1 mice had developed AD and had developed deficits in spatial memory. Furthermore, the escape latency of the experimental group mice was significantly decreased compared to the pathological control group, even approaching that of the normal control group mice. This experimental result demonstrates that the Chinese herbal composition of the present invention can effectively treat AD and improve spatial memory in mice with AD.

[0104] D. Immunofluorescence staining analysis:

[0105] Figure 6 shows the number of Aβ plaques measured in the brains of mice in each group. As shown in Figure 6, compared to the normal control group, the number of Aβ plaques in the pathological control group mice increased significantly, indicating that the APP / PS1 mice had developed AD and had abnormally accumulated a large number of Aβ plaques in their brains. Furthermore, compared to the pathological control group, the number of Aβ plaques in the experimental group mice decreased significantly. These experimental results demonstrate that the traditional Chinese medicine composition of the present invention can effectively treat AD and improve Aβ plaque accumulation.

[0106] E. Enzyme-binding immunosorbent assay:

[0107] Figure 7 shows the soluble Aβ40 levels measured in the cerebral cortex of each group of mice. As shown in Figure 7, compared to the normal control group, the soluble Aβ40 levels in the pathological control group mice were significantly increased, indicating that the APP / PS1 mice had developed AD and produced a large amount of soluble Aβ40 in the brain. Furthermore, compared to the pathological control group, the soluble Aβ40 levels in the experimental group mice were significantly decreased. These experimental results demonstrate that the traditional Chinese medicine composition of the present invention can effectively treat AD and reduce the production of soluble Aβ40.

[0108] Example 5. Effect of the dosage of the Chinese medicine composition of the present invention on the therapeutic effect of mice with AD

[0109] Experimental methods:

[0110] A. Administration of the Chinese medicine composition of the present invention:

[0111] First, APP / PS1 mice (4 months old) were randomly divided into 1 pathological control group and 2 experimental groups (i.e., experimental groups 1 and 2), and C57 / BL6 mice (4 months old) were used as a normal control group, with n = 5 per group. By oral gavage, the mice in experimental groups 1 and 2 were respectively administered with the Chinese medicine composition 3 prepared in Example 1 (doses of 50 and 300 mg / kg, respectively, both in normal saline), while the pathological control group and the normal control group were administered with an equal amount of normal saline, once a day, for a total of 18 weeks. At the end of the 14th week after the start of administration, the mice in each group were taken for the test in item B below, and at the end of the 18th week after the start of administration, the mice in each group were sacrificed and the brains were removed, and then taken for the analysis in item C below.

[0112] B. Morse water maze test:

[0113] This test was carried out according to the method described in Example 4, item D above.

[0114] C. Immunofluorescence staining analysis:

[0115] First, tissue slices were prepared from the brains of each group of mice according to the method described in Item E of Example 4 above. Then, some of the obtained brain tissue slices were selected and subjected to immunofluorescence staining analysis and calculation of the number of Aβ plaques according to the method described in Item E of Example 4 above.

[0116] In addition, the hippocampus tissue sections were selected from the obtained brain tissue sections, and then anti-doublecortin (DCX) antibody (Abcam, Cat#ab18723) was used as the primary antibody and Alexa- 647-conjugated anti-IgG antibody (Jackson ImmunoResearch) was used as the secondary antibody and immunofluorescence staining was performed according to the techniques well known and commonly used by those skilled in the art. The stained tissue sections were observed and photographed using a conjugate fluorescence microscope, and then the DCX in the subgranular zone of the dentate gyrus was counted using Image J software. + The number of cells.

[0117] The obtained experimental data were then analyzed according to the method described in item 1 of the “General Experimental Methods” above.

[0118] result:

[0119] A. Morse water maze test:

[0120] Figure 8 shows the escape latency measured for each group of mice. As shown in Figure 8 , the escape latency of mice in the pathological control group was significantly increased compared to the normal control group, indicating that the APP / PS1 mice had developed AD and had developed deficits in spatial memory. Furthermore, the escape latency of mice in experimental groups 1 and 2 was significantly decreased compared to the pathological control group, even approaching that of mice in the normal control group. These experimental results demonstrate that the Chinese herbal composition of the present invention, at various doses, is effective in treating AD and improving spatial memory in mice with AD.

[0121] B. Immunofluorescence staining analysis:

[0122] Figure 9 shows the number of Aβ plaques measured in the brains of mice in each group. As shown in Figure 9, compared to the normal control group, the number of Aβ plaques in the pathological control group mice increased significantly, indicating that the APP / PS1 mice had developed AD and had abnormally accumulated a large number of Aβ plaques in their brains. Furthermore, compared to the pathological control group, the number of Aβ plaques in mice in experimental groups 1 and 2 decreased significantly.

[0123] Figure 10 shows the DCX measured in the subgranular area of ​​the dentate gyrus of the brain of each group of mice + As shown in Figure 10, compared with the normal control group, the DCX + There was a significant decrease in the number of cells, which indicated that APP / PS1 mice had developed AD and inhibited neurogenesis in the hippocampus. In addition, compared with the pathological control group, the DCX + The cell numbers increased significantly.

[0124] These experimental results show that the Chinese medicine composition of the present invention can improve Aβ plaque aggregation and promote neurogenesis at different doses, thereby achieving the effect of treating AD.

[0125] Example 6. Evaluation of the efficacy of the Chinese herbal composition of the present invention in improving AD patients

[0126] A. Preparation of Chinese medicine composition 4 of the present invention:

[0127] The six powdered ingredients were mixed according to Table 3 below to obtain the Chinese medicine composition 4 of the present invention.

[0128] Table 3. Formula of Chinese medicine composition 4 of the present invention

[0129] B. Experimental methods and results:

[0130] The subjects participating in this trial were all from the Tri-Service General Hospital and were diagnosed with mild to moderate AD through outpatient clinics. A total of 42 participants participated.

[0131] All subjects were randomly divided into an experimental group (n = 25) and a pathological control group (n = 17). The subjects in the experimental group were orally administered the Chinese medicine composition 4 prepared in item A of this example (dose of 6 g) twice a day for a total of 3 months. The subjects were scored using the Mini-Mental State Examination (MMSE) before and after the start of the administration.

[0132] The obtained results are shown in Table 4 below.

[0133] Table 4. MMSE scores of subjects *: Compared with the pathological control group, p < 0.05

[0134] As shown in Table 4, after 3 months of administration, the MMSE scores of the experimental group subjects increased significantly, while those in the pathological control group showed a slight decrease. This experimental result shows that the Chinese herbal composition of the present invention can effectively treat AD and improve cognitive function. Example 7. Evaluation of the therapeutic effect of the Chinese herbal composition of the present invention on mice with ischemia-reperfusion-induced nerve injury

[0135] Experimental methods:

[0136] A. Induction of nerve damage and administration of the Chinese medicine composition of the present invention:

[0137] First, ICR mice were randomly divided into one normal control group, one pathological control group, and one experimental group, with n = 5 per group. Then, the pathological control and experimental group mice were anesthetized with a mixture of 1.5-2% isoflurane and oxygen. A midline cervical incision was performed to expose the common, external, and internal carotid arteries. The following middle cerebral artery occlusion / reperfusion (MCAO / R) procedure was performed: a heat-blunted nylon monofilament surgical suture (approximately 100 μm in diameter) coated with silicone was placed into the exposed external carotid artery and advanced to the internal carotid artery and into the circle of Willis, thereby occluding the right middle cerebral artery (MCA) and blocking blood flow. After 40 minutes, the surgical sutures were withdrawn, causing reperfusion of the middle cerebral artery, thereby causing the mice to suffer from ischemia-reperfusion-induced neurological damage. Mice in the normal control group received no treatment.

[0138] Two hours after MCAO / R, the experimental group mice were administered the Chinese medicine composition 3 prepared in Example 1 (at a dose of 1 g / kg, in normal saline) via oral gavage. The pathological control group and the normal control group mice were administered an equal amount of normal saline once daily until the second day after MCAO / R.

[0139] On days 0, 1, and 2 after MCAO / R, the number of surviving mice in each group was observed. The survival rate (%) of each group of mice was calculated by substituting the obtained number of surviving mice into the following formula (2):

[0140] Formula (2): D = (E / 5) × 100%

[0141] Where: D = survival rate (%)

[0142] E = number of survivors in each group

[0143] In addition, on the first day after MCAO / R, mice in the pathology control group and the experimental group were used for analysis in item B below. On the second day after MCAO / R and after the completion of the survival rate analysis, the mice in each group were sacrificed and the brains were removed, and then used for analysis in item C below.

[0144] B. Analysis of mobility:

[0145] The mice in the pathological control group and the experimental group were placed in a behavioral observation box (60 x 40 x 60 cm 3 ) and used a video-tracking system (SMART v2.5.21, Panlab, Spain) to analyze the movement distance and movement area of ​​each mouse within 3 minutes.

[0146] The obtained experimental data were then analyzed according to the method described in item 1 of the “General Experimental Methods” above.

[0147] C. Immunofluorescence staining analysis:

[0148] The brains of mice in each group were fixed with 4% formaldehyde at 4°C and then sectioned (approximately 20 μm thick) using a freezing microtome. The resulting tissue sections were immunofluorescently stained using the primary and secondary antibodies listed in Table 5 below, according to techniques well known and commonly used by those skilled in the art, and then mounted with an anti-fluorescence-fading mounting medium containing the nuclear stain DAPI. The stained tissue sections were observed and photographed using a confocal fluorescence microscope (Zeiss LSM 780). The number of neuronal stem cells in the hippocampus and lateral ventricle was then counted using Zen 2011 imaging software (black edition, Carl Zeiss MicroImaging GmbH, 1997-2011).

[0149] The obtained experimental data were then analyzed according to the method described in item 1 of the “General Experimental Methods” above.

[0150] Table 5. Primary and secondary antibodies used for immunofluorescence staining analysis

[0151] result:

[0152] A. Analysis of survival rate:

[0153] Table 6 below shows the survival rates of mice in each group. As shown in Table 6, the survival rate of mice in the pathological control group decreased rapidly and significantly compared to the normal control group, indicating that MCAO / R successfully inflicted ischemia-reperfusion-induced neurological damage in the mice. Furthermore, the survival rate of mice in the experimental group increased significantly compared to the pathological control group.

[0154] Table 6. Survival rate of mice in each group (%)

[0155] B. Analysis of mobility:

[0156] Table 7 below shows the measured movement distances and movement areas of the mice in each group. As can be seen from Table 7, the movement distances and movement areas of the mice in the experimental group were significantly greater than those in the pathological control group.

[0157] Table 7. Movement distance and movement area measured for each group of mice *: Compared with the pathological control group, p < 0.05

[0158] C. Immunofluorescence staining analysis:

[0159] Figure 11 shows the immunofluorescence staining analysis results of the hippocampus of each group of mice. Table 8 below shows the number of neuronal stem cells measured in the hippocampus and lateral ventricle of each group of mice.

[0160] Table 8. The number of neuronal stem cells measured in the hippocampus and lateral ventricle of the brain of each group of mice *: Compared with the pathological control group, p < 0.05

[0161] As shown in Figure 11 and Table 8, compared to the normal control group, the pathological control group mice showed a significant decrease in both neuronal stem cells and microglia, indicating that MCAO / R successfully inflicted ischemia-reperfusion-induced neurological damage in the mice. Furthermore, compared to the pathological control group, the experimental group mice showed significant proliferation of neuronal stem cells and microglia. In particular, the number of neuronal stem cells in the lateral ventricles was even close to that of the normal control group.

[0162] These experimental results show that the Chinese medicine composition of the present invention can effectively treat ischemia-reperfusion-induced nerve damage and promote neurogenesis to repair damaged brain areas, thereby improving mobility and survival ability.

[0163] Based on the above experimental results, the applicant believes that the Chinese medicine composition of the present invention can be used to treat dementia, including degenerative dementia, vascular dementia and mixed dementia, especially AD.

[0164] All patents and publications cited in this specification are hereby incorporated by reference in their entirety. In the event of any conflict, the detailed description of this specification (including definitions) will prevail.

[0165] Although the present invention has been described with reference to the specific embodiments above, it is apparent that many modifications and variations can be made without departing from the scope and spirit of the invention. It is therefore intended that the present invention be limited only as indicated by the appended claims.

Claims

1. A Chinese medicine composition, characterized in that: The traditional Chinese medicine composition comprises: red astragalus root, scutellaria root, asparagus root and ligusticum chuanxiong.

2. The Chinese medicine composition according to claim 1, characterized in that: The traditional Chinese medicine composition comprises 7-11 parts by weight of red astragalus root, 2-6 parts by weight of scutellaria root, 4-7 parts by weight of asparagus root and 2-4 parts by weight of ligusticum chuanxiong.

3. The Chinese medicine composition according to claim 2, characterized in that: The traditional Chinese medicine composition comprises 8-10 parts by weight of red astragalus root, 3-5 parts by weight of scutellaria root, 5-6 parts by weight of asparagus root and 3 parts by weight of ligusticum chuanxiong.

4. The Chinese medicine composition according to claim 1, characterized in that: The traditional Chinese medicine composition further comprises deer antler.

5. The Chinese medicine composition according to claim 4, characterized in that: The traditional Chinese medicine composition comprises 7-11 parts by weight of red astragalus root, 2-6 parts by weight of scutellaria root, 4-7 parts by weight of asparagus root, 2-4 parts by weight of ligusticum chuanxiong and 1-3 parts by weight of pilose antler.

6. The Chinese medicine composition according to claim 5, characterized in that: The traditional Chinese medicine composition comprises 8-10 parts by weight of red astragalus root, 3-5 parts by weight of scutellaria root, 5-6 parts by weight of asparagus root, 3 parts by weight of chuanxiong rhizome and 2 parts by weight of pilose antler.

7. The Chinese medicine composition according to claim 4, characterized in that: The traditional Chinese medicine composition further comprises aconite root.

8. The Chinese medicine composition according to claim 7, characterized in that: The traditional Chinese medicine composition comprises 7-11 parts by weight of red astragalus root, 2-6 parts by weight of scutellaria root, 4-7 parts by weight of asparagus root, 2-4 parts by weight of chuanxiong rhizome, 1-3 parts by weight of pilose antler and 1-3 parts by weight of aconite root.

9. The Chinese medicine composition according to claim 8, characterized in that: The traditional Chinese medicine composition comprises 8-10 parts by weight of red astragalus root, 3-5 parts by weight of scutellaria baicalensis root, 5-6 parts by weight of asparagus root, 3 parts by weight of chuanxiong rhizome, 2 parts by weight of pilose antler and 2 parts by weight of aconite root.

10. A Chinese medicine composition for use in preparing a medicine for treating dementia, characterized in that: The traditional Chinese medicine composition comprises: red astragalus root, scutellaria root, asparagus root and ligusticum chuanxiong.

11. The use according to claim 10, characterized in that: The traditional Chinese medicine composition comprises 7-11 parts by weight of red astragalus root, 2-6 parts by weight of scutellaria root, 4-7 parts by weight of asparagus root and 2-4 parts by weight of ligusticum chuanxiong.

12. The use according to claim 11, characterized in that: The traditional Chinese medicine composition comprises 8-10 parts by weight of red astragalus root, 3-5 parts by weight of scutellaria root, 5-6 parts by weight of asparagus root and 3 parts by weight of ligusticum chuanxiong.

13. The use according to claim 10, characterized in that: The traditional Chinese medicine composition further comprises deer antler.

14. The use according to claim 13, characterized in that: The traditional Chinese medicine composition comprises 7-11 parts by weight of red astragalus root, 2-6 parts by weight of scutellaria root, 4-7 parts by weight of asparagus root, 2-4 parts by weight of ligusticum chuanxiong and 1-3 parts by weight of pilose antler.

15. The use according to claim 14, characterized in that: The traditional Chinese medicine composition comprises 8-10 parts by weight of red astragalus root, 3-5 parts by weight of scutellaria root, 5-6 parts by weight of asparagus root, 3 parts by weight of chuanxiong rhizome and 2 parts by weight of pilose antler.

16. The use according to claim 13, characterized in that: The traditional Chinese medicine composition further comprises aconite root.

17. The use according to claim 16, characterized in that: The traditional Chinese medicine composition comprises 7-11 parts by weight of red astragalus root, 2-6 parts by weight of scutellaria root, 4-7 parts by weight of asparagus root, 2-4 parts by weight of chuanxiong rhizome, 1-3 parts by weight of pilose antler and 1-3 parts by weight of aconite root.

18. The use according to claim 17, characterized in that: The traditional Chinese medicine composition comprises 8-10 parts by weight of red astragalus root, 3-5 parts by weight of scutellaria baicalensis root, 5-6 parts by weight of asparagus root, 3 parts by weight of chuanxiong rhizome, 2 parts by weight of pilose antler and 2 parts by weight of aconite root.

19. The use according to claim 10, characterized in that: The dementia is selected from the group consisting of degenerative dementia, vascular dementia, and mixed dementia.

20. The use according to claim 19, characterized in that: The dementia is a degenerative dementia selected from the group consisting of Alzheimer's disease, Lewy body dementia, frontotemporal dementia, Parkinson's disease dementia, and Huntington's disease.

21. The use according to claim 20, characterized in that: The dementia is Alzheimer's disease.

22. The use according to claim 10, characterized in that: Treatment for this dementia involves promoting neurogenesis.

23. A method for treating dementia, comprising administering a Chinese medicine composition to a subject in need thereof, wherein: The traditional Chinese medicine composition comprises: red astragalus root, scutellaria root, asparagus root and ligusticum chuanxiong.

24. The method according to claim 23, wherein: The traditional Chinese medicine composition comprises 7-11 parts by weight of red astragalus root, 2-6 parts by weight of scutellaria root, 4-7 parts by weight of asparagus root and 2-4 parts by weight of ligusticum chuanxiong.

25. The method according to claim 24, wherein: The traditional Chinese medicine composition comprises 8-10 parts by weight of red astragalus root, 3-5 parts by weight of scutellaria root, 5-6 parts by weight of asparagus root and 3 parts by weight of ligusticum chuanxiong.

26. The method according to claim 23, wherein: The traditional Chinese medicine composition further comprises deer antler.

27. The method according to claim 26, wherein: The traditional Chinese medicine composition comprises 7-11 parts by weight of red astragalus root, 2-6 parts by weight of scutellaria root, 4-7 parts by weight of asparagus root, 2-4 parts by weight of ligusticum chuanxiong and 1-3 parts by weight of pilose antler.

28. The method according to claim 27, wherein: The traditional Chinese medicine composition comprises 8-10 parts by weight of red astragalus root, 3-5 parts by weight of scutellaria root, 5-6 parts by weight of asparagus root, 3 parts by weight of chuanxiong rhizome and 2 parts by weight of pilose antler.

29. The method according to claim 26, wherein: The traditional Chinese medicine composition further comprises aconite root.

30. The method according to claim 29, wherein: The traditional Chinese medicine composition comprises 7-11 parts by weight of red astragalus root, 2-6 parts by weight of scutellaria root, 4-7 parts by weight of asparagus root, 2-4 parts by weight of chuanxiong rhizome, 1-3 parts by weight of pilose antler and 1-3 parts by weight of aconite root.

31. The method according to claim 30, wherein: The traditional Chinese medicine composition comprises 8-10 parts by weight of red astragalus root, 3-5 parts by weight of scutellaria baicalensis root, 5-6 parts by weight of asparagus root, 3 parts by weight of chuanxiong rhizome, 2 parts by weight of pilose antler and 2 parts by weight of aconite root.

32. The method according to claim 23, wherein: The dementia is selected from the group consisting of degenerative dementia, vascular dementia, and mixed dementia.

33. The method according to claim 32, wherein: The dementia is a degenerative dementia selected from the group consisting of Alzheimer's disease, Lewy body dementia, frontotemporal dementia, Parkinson's disease dementia, and Huntington's disease.

34. The method according to claim 33, wherein: The dementia is Alzheimer's disease.

35. The method according to claim 23, wherein: Treatment for this dementia involves promoting neurogenesis.

36. A Chinese medicine composition for treating dementia, characterized by: The traditional Chinese medicine composition comprises: red astragalus root, scutellaria root, asparagus root and ligusticum chuanxiong.

37. The Chinese medicine composition according to claim 36, characterized in that: The traditional Chinese medicine composition comprises 7-11 parts by weight of red astragalus root, 2-6 parts by weight of scutellaria root, 4-7 parts by weight of asparagus root and 2-4 parts by weight of ligusticum chuanxiong.

38. The Chinese medicine composition according to claim 37, characterized in that: The traditional Chinese medicine composition comprises 8-10 parts by weight of red astragalus root, 3-5 parts by weight of scutellaria root, 5-6 parts by weight of asparagus root and 3 parts by weight of ligusticum chuanxiong.

39. The Chinese medicine composition according to claim 36, characterized in that: The traditional Chinese medicine composition further comprises deer antler.

40. The Chinese medicine composition according to claim 39, characterized in that: The traditional Chinese medicine composition comprises 7-11 parts by weight of red astragalus root, 2-6 parts by weight of scutellaria root, 4-7 parts by weight of asparagus root, 2-4 parts by weight of ligusticum chuanxiong and 1-3 parts by weight of pilose antler.

41. The Chinese medicine composition according to claim 40, characterized in that: The traditional Chinese medicine composition comprises 8-10 parts by weight of red astragalus root, 3-5 parts by weight of scutellaria root, 5-6 parts by weight of asparagus root, 3 parts by weight of chuanxiong rhizome and 2 parts by weight of pilose antler.

42. The Chinese medicine composition according to claim 39, characterized in that: The traditional Chinese medicine composition further comprises aconite root.

43. The Chinese medicine composition according to claim 42, characterized in that: The traditional Chinese medicine composition comprises 7-11 parts by weight of red astragalus root, 2-6 parts by weight of scutellaria root, 4-7 parts by weight of asparagus root, 2-4 parts by weight of chuanxiong rhizome, 1-3 parts by weight of pilose antler and 1-3 parts by weight of aconite root.

44. The Chinese medicine composition according to claim 43, characterized in that: The traditional Chinese medicine composition comprises 8-10 parts by weight of red astragalus root, 3-5 parts by weight of scutellaria baicalensis root, 5-6 parts by weight of asparagus root, 3 parts by weight of chuanxiong rhizome, 2 parts by weight of pilose antler and 2 parts by weight of aconite root.

45. The Chinese medicine composition according to claim 36, characterized in that: The dementia is selected from the group consisting of degenerative dementia, vascular dementia, and mixed dementia.

46. ​​The Chinese medicine composition according to claim 45, characterized in that: The dementia is a degenerative dementia selected from the group consisting of Alzheimer's disease, Lewy body dementia, frontotemporal dementia, Parkinson's disease dementia, and Huntington's disease.

47. The Chinese medicine composition according to claim 46, characterized in that: The dementia is Alzheimer's disease.

48. The Chinese medicine composition according to claim 36, characterized in that: Treatment for this dementia involves promoting neurogenesis.

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